Epigenetic targeting therapy for tumor radiosensitization

Introduction: A malignant tumor is composed of a hierarchically organized, heterogeneous pool of cells in various stages of differentiation including cancer stem cells (CSCs) as the main cell population responsible for tumor initiation, growth and relapse as well as for metastasis formation and therapy resistance. Preliminary work of our group showed that photon irradiation of prostate cancer cells induces an augmentation of the CSC population that can be attributed to an epigenetic reprogramming of non-CSCs into CSCs. The combination of the histone methyltransferase EZH2 inhibitor 3-Deazaneplanocin A (DZNep) with x-ray irradiation leads to a radiosensitization and prevention of the cellular reprogramming of prostate cancer cells in vitro, in vivo and in ex vivo treated primary prostate cancer samples.

Objectives: The aim of this study is (1) the investigation of potential radiosensitizing effects of epigenetic inhibitors for different tumor entities including prostate cancer, head and neck squamous cell carcinoma (HNSCC) and glioblastoma multiforme (GBM); (2) a high throughput screening (HTS) using a chemical library with epigenetic targeting substances to identify novel epigenetic targeting agents for tumor radiosensitization and (3) the correlation of radiation-induced CSC and epigenetic markers after photon versus 150MeV proton irradiation to identify biomarkers for a personalized proton therapy.

Methods: Evaluation of the cytotoxicity (MTT-Assay, CellTiterGlo®-Assay), radiosensitivity (colony formation assay), DNA repair capacity (γH2AX foci assay), CSC marker expression (Flow cytometry) and histone modifications (Western blot) for prostate cancer (DU145, PC3), HNSCC (FaDu, Cal33) and GBM (LN229, U87MG) cell lines using different clinically relevant epigenetic modulators in combination with irradiation.

Results: Our results show that epigenetic marks including histone modifications are modulated after ionizing radiation in the different tumor entities including prostate cancer, HNSCC and GBM and that treatment with some epigenetic inhibitors leads to the radiosensitization of cancer cells. Furthermore, we found differences in CSC marker expression and epigenetic modulation after photon versus proton irradiation.

Conclusion: Epigenetic targeting therapy may be useful as a co-therapy strategy to prevent tumor cell reprogramming and promote radiosensitization of the different tumor entities. Moreover, CSC and histone mark-based biomarkers can be potentially used as predictive markers for a personalized radiotherapy.